Human embryonic stem cells (hESCs) hold significant promise for regenerative medicine. In this application our goal is to derive hESC lines from pre-implantation embryos to generate a source of low passage lines that can be used in research and to develop the procedures required to generate a clinic grade cell-based product. In this application we will develop the procedures for deriving, expanding and banking using current good manufacturing practices (cGMP), which are necessary for producing a clinic grade line. We aim to do this using chemically defined media and surfaces in consultation with the Director of Regulatory Compliance at our Institution. Secondly, we aim to derive hESC lines from embryos that have genetic disorders. These genetic aberrations are identified following pre implantation genetic diagnosis (PGD) or pre implantation genetic screening (PGS) of consented embryos from couples undergoing in vitro fertilization (IVF). Deriving hESC lines from these embryos will be essential for understanding the underlying causes of spontaneous miscarriage, or as a tool to improve the quality of life of individuals born with these chromosomal abnormalities including Trisomy 21 (Down Syndrome), or Monosomy X (Turner Syndrome).

Statement of Benefit to California:

Human embryonic stem cells could revolutionize modern medicine if used in cell-based therapies. However, the translational use of hESCs will not be realized unless an appropriate repository of low passage cells is derived and made available. This resource will solidify California as a recognized world leader in ESC research through the generation of a centralized and comprehensive resource of hESC lines and the creation of jobs to support this effort, and validate the decision of California voters who ensured the passage of Proposition 71. Our proposal works toward generating lines that make treatment strategies from hESCs one-step closer.

Progress Report:

Year 1

In the first year of funding, we have made excellent progress towards our goal of deriving pluripotent stem cell lines from human embryos. We have generated twenty new human embryonic stem cell lines in total, with eight of these new lines carrying a genetic abnormality identified by pre-implantation genetic diagnosis (PGD) or pre-implantation genetic screening (PGS) in the embryo, which was subsequently confirmed in the established embryonic stem cell line. These aneuploid lines represent new cell-based models of human disease. We have also confirmed the fidelity of a feeder-free system with which to derive new pluripotent stem cell lines towards the generation of standard operating procedures for current good manufacturing practice of embryonic stem cell derivation and maintenance.

Year 2

The goal of this CIRM New Cell Line award is to derive new pluripotent stem cell lines from human embryos for research, and also to develop the conditions for deriving embryonic stem cell lines by current good manufacturing practice. In this reporting period we are documenting the derivation of twenty one new embryonic stem cell lines. Nineteen lines have been Karyotyped, and twelve out of nineteen are diagnosed as euploid with a karyotype of either 46, XX or 46, XY. Three of the karyotypically normal cell lines (UCLA-1, UCLA-2 and UCLA-3), have been registered in the NIH stem cell registry as well as registered at CIRM. One of the aneuploid cell lines represents a new disease model for Trisomy 21.

Year 3

This is a new cell lines award which involves the derivation of new human embryonic stem cell lines from human blastocysts. In this reporting period we have generated fifteen new hESC lines. In particular, we have generated two new disease hESC lines for Patau syndrome (Trisomy 31) and an additional disease hESC line for Down Syndrome (Trisomy 21). We have also generated a comprehensive collection of standard operating procedures (SOPs) for the current good manufacturing of new hESC lines at our institution.

Year 4

In the no cost extension period for this new cell line award we completed the basic analysis of ten human embryonic stem cell (hESC) lines generated at UCLA and performed a comprehensive analysis of five additional hESC lines with trisomy 21 which constitute a unique disease model for Down Syndrome. Our data on the five Down Syndrome hESC lines reveals unique abnormalities in neural progenitor cell differentiation. Taken together, this New Cell Line Award has resulted in the generation of unique hESC tools for the scientific community, and in particular we demonstrate that hESC lines with Trisomy 21 have the potential to contribute unique perspectives on the underlying mechanisms of disease in individuals with Down Syndrome.